title: Isolation, culture, and functional characterization of adult mouse cardiomyoctyes.
abstract: The use of primary cardiomyocytes (CMs) in culture has provided a powerful complement to murine models of heart disease in advancing our understanding of heart disease. In particular, the ability to study ion homeostasis, ion channel function, cellular excitability and excitation-contraction coupling and their alterations in diseased conditions and by disease-causing mutations have led to significant insights into cardiac diseases. Furthermore, the lack of an adequate immortalized cell line to mimic adult CMs, and the limitations of neonatal CMs (which lack many of the structural and functional biomechanics characteristic of adult CMs) in culture have hampered our understanding of the complex interplay between signaling pathways, ion channels and contractile properties in the adult heart strengthening the importance of studying adult isolated cardiomyocytes. Here, we present methods for the isolation, culture, manipulation of gene expression by adenoviral-expressed proteins, and subsequent functional analysis of cardiomyocytes from the adult mouse. The use of these techniques will help to develop mechanistic insight into signaling pathways that regulate cellular excitability, Ca(2+) dynamics and contractility and provide a much more physiologically relevant characterization of cardiovascular disease.
authors.affiliation: Cardiovascular Institute, Beth Israel Deaconess Medical Center, Harvard Medical School.
authors.forename: Evan Lee
authors.initials: EL
authors.lastname: Graham
authors.forename: Cristina
authors.initials: C
authors.lastname: Balla
authors.forename: Hannabeth
authors.initials: H
authors.lastname: Franchino
authors.forename: Yonathan
authors.initials: Y
authors.lastname: Melman
authors.forename: Federica
authors.initials: F
authors.lastname: del Monte
authors.forename: Saumya
authors.initials: S
authors.lastname: Das
mesh_terms: D000818:Animals; D003584:Cytological Techniques / Q000379:methods*; D051379:Mice; D032383:Myocytes, Cardiac / Q000166:cytology* / Q000502:physiology*; D018408:Patch-Clamp Techniques; D014162:Transfection